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Multivalent Rab interactions determine tether-mediated membrane fusion

Membrane fusion at endomembranes requires cross-talk between Rab GTPases and tethers to drive SNARE-mediated lipid bilayer mixing. Several tethers have multiple Rab-binding sites with largely untested function. Here we dissected the lysosomal HOPS complex as a tethering complex with just two binding...

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Autores principales: Lürick, Anna, Gao, Jieqiong, Kuhlee, Anne, Yavavli, Erdal, Langemeyer, Lars, Perz, Angela, Raunser, Stefan, Ungermann, Christian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5231900/
https://www.ncbi.nlm.nih.gov/pubmed/27852901
http://dx.doi.org/10.1091/mbc.E16-11-0764
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author Lürick, Anna
Gao, Jieqiong
Kuhlee, Anne
Yavavli, Erdal
Langemeyer, Lars
Perz, Angela
Raunser, Stefan
Ungermann, Christian
author_facet Lürick, Anna
Gao, Jieqiong
Kuhlee, Anne
Yavavli, Erdal
Langemeyer, Lars
Perz, Angela
Raunser, Stefan
Ungermann, Christian
author_sort Lürick, Anna
collection PubMed
description Membrane fusion at endomembranes requires cross-talk between Rab GTPases and tethers to drive SNARE-mediated lipid bilayer mixing. Several tethers have multiple Rab-binding sites with largely untested function. Here we dissected the lysosomal HOPS complex as a tethering complex with just two binding sites for the Rab7-like Ypt7 protein to determine their relevance for fusion. Using tethering and fusion assays combined with HOPS mutants, we show that HOPS-dependent fusion requires both Rab-binding sites, with Vps39 being the stronger Ypt7 interactor than Vps41. The intrinsic amphipathic lipid packaging sensor (ALPS) motif within HOPS Vps41, a target of the vacuolar kinase Yck3, is dispensable for tethering and fusion but can affect tethering if phosphorylated. In combination, our data demonstrate that a multivalent tethering complex uses its two Rab bindings to determine the place of SNARE assembly and thus fusion at endomembranes.
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spelling pubmed-52319002017-03-30 Multivalent Rab interactions determine tether-mediated membrane fusion Lürick, Anna Gao, Jieqiong Kuhlee, Anne Yavavli, Erdal Langemeyer, Lars Perz, Angela Raunser, Stefan Ungermann, Christian Mol Biol Cell Articles Membrane fusion at endomembranes requires cross-talk between Rab GTPases and tethers to drive SNARE-mediated lipid bilayer mixing. Several tethers have multiple Rab-binding sites with largely untested function. Here we dissected the lysosomal HOPS complex as a tethering complex with just two binding sites for the Rab7-like Ypt7 protein to determine their relevance for fusion. Using tethering and fusion assays combined with HOPS mutants, we show that HOPS-dependent fusion requires both Rab-binding sites, with Vps39 being the stronger Ypt7 interactor than Vps41. The intrinsic amphipathic lipid packaging sensor (ALPS) motif within HOPS Vps41, a target of the vacuolar kinase Yck3, is dispensable for tethering and fusion but can affect tethering if phosphorylated. In combination, our data demonstrate that a multivalent tethering complex uses its two Rab bindings to determine the place of SNARE assembly and thus fusion at endomembranes. The American Society for Cell Biology 2017-01-15 /pmc/articles/PMC5231900/ /pubmed/27852901 http://dx.doi.org/10.1091/mbc.E16-11-0764 Text en © 2017 Lürick et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.
spellingShingle Articles
Lürick, Anna
Gao, Jieqiong
Kuhlee, Anne
Yavavli, Erdal
Langemeyer, Lars
Perz, Angela
Raunser, Stefan
Ungermann, Christian
Multivalent Rab interactions determine tether-mediated membrane fusion
title Multivalent Rab interactions determine tether-mediated membrane fusion
title_full Multivalent Rab interactions determine tether-mediated membrane fusion
title_fullStr Multivalent Rab interactions determine tether-mediated membrane fusion
title_full_unstemmed Multivalent Rab interactions determine tether-mediated membrane fusion
title_short Multivalent Rab interactions determine tether-mediated membrane fusion
title_sort multivalent rab interactions determine tether-mediated membrane fusion
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5231900/
https://www.ncbi.nlm.nih.gov/pubmed/27852901
http://dx.doi.org/10.1091/mbc.E16-11-0764
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